Characterization of Recombinant Endonuclease IV from Mycobacterium tuberculosis

M. A. Dymova, A. V. Endutkin, V. V. Polunovsky, A. I. Zakabunin, E. A. Khrapov, N. A. Torgasheva, A. V. Yudkina, G. V. Mechetin, M. L. Filipenko, D. O. Zharkov

Research output: Contribution to journalArticlepeer-review

Abstract

Mycobacterium tuberculosis cells contain two apurinic/apyrimidinic (AP) endonucleases, endonuclease IV (MtbEnd) and exonuclease III (MtbXthA), the former playing a dominant role in protecting mycobacterial DNA from oxidative stress. Mycobacterial endonuclease IV substantially differs from its homologs found in Escherichia coli and other proteobacteria in a number of conserved positions important for DNA binding and AP site recognition. The M. tuberculosis end gene was cloned, and recombinant MtbEnd purified and characterized. The protein efficiently hydrolyzed DNA at the natural AP site and its 1′-deoxy analog in the presence of divalent cations, of which Ca2+, Mn2+, and Co2+ supported the highest activity. Exonuclease activity was not detected in MtbEnt preparations. The pH optimum was estimated at 7.0–8.0; the ionic strength optimum, at ~50 mM NaCl. Enzymatic activity of MtbEnd was suppressed in the presence of methoxyamine, a chemotherapeutic agent that modifies AP sites. Based on the results, MtbEnd was assumed to provide a possible target for new anti-tuberculosis drugs.

Original languageEnglish
Pages (from-to)225-233
Number of pages9
JournalMolecular Biology
Volume55
Issue number2
DOIs
Publication statusPublished - Mar 2021

Keywords

  • AP endonuclease
  • DNA repair
  • methoxyamine
  • Mycobacterium tuberculosis

OECD FOS+WOS

  • 1.06 BIOLOGICAL SCIENCES

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